蛋白酶激活受体-1拮抗剂对矽肺大鼠肺纤维化的干预作用  

Intervention effect of protease activated receptor-1 antagonist on pulmonary fibrosis in silicosis rats

作　　者：刘潇蔓 杨碧珺 厉铭 李超[1] 薄存香[1] 孟祥敬 贾强[1,2] LIU Xiaoman;YANG Bijun;LI Ming;LI Chao;BO Cunxiang;MENG Xiangjing;JIA Qiang(Shandong Provincial Academy of Occupational Health and Occupational Medicine,Shandong First Medical University/Shandong Provincial Academy of Medical Sciences,Jinan,Shandong 250062,China;不详)

机构地区：[1]山东省职业卫生与职业病防治研究院,山东第一医科大学/山东省医学科学院,山东济南250062 [2]山东第一医科大学公共卫生与健康管理学院/山东省医学科学院

出　　处：《中国工业医学杂志》2025年第1期5-9,60,I0004,共7页Chinese Journal of Industrial Medicine

基　　金：“十四五”国家重点研发计划项目(2022YFC2503202);山东省医药卫生科技项目(202312010678);山东省博士后基金(SDCX-ZG-202203045);山东省自然科学基金(ZR2022MH293);济南市科技局临床医学科技创新计划(202328075,202430001)。

摘　　要：目的 探讨矽肺大鼠凝血级联反应变化情况以及蛋白酶激活受体-1(PAR-1)拮抗剂对其肺纤维化的干预作用。方法 27只SPF级健康雄性Wistar大鼠随机分为对照组、模型组(SiO_(2))及Vorapaxar(PAR-1拮抗剂)组,采用口鼻吸入暴露法建立矽肺大鼠模型;SiO_(2)粉尘浓度800 mg/m^(3),染尘6 h/d,连续吸入28 d;Vorapaxar组动物于造模第1天开始给予Vorapaxar(3.5 mg/kg)灌胃。每组动物分别在染毒后第14、28、56 d处死,收集肺组织及血清;采用HE、Masson染色观察各组大鼠肺组织病理和纤维化变化,免疫组化法检测肺组织转化生长因子(TGF)-β1表达,酶联免疫吸附试验(ELISA)检测各组大鼠血清克拉拉蛋白(CC16)、组织因子(TF)、凝血酶-抗凝血酶复合物(TAT)和纤溶酶原激活物抑制因子(PAI-1)水平,蛋白免疫印迹试验(Western blot)检测各组大鼠肺组织胶原蛋白Ⅰ(Col-Ⅰ)、整合素(αvβ)5、αvβ6、PAR-1和结缔组织生长因子(CTGF)的蛋白表达。结果 染尘14、28、56 d时,血清中TF、TAT和PAI-1水平均较对照组明显升高(P<0.05),但CC16水平显著降低(P<0.05);模型组大鼠染尘56 d时肺内炎症和纤维化程度,肺组织Col-Ⅰ、αvβ5、αvβ6、PAR-1、CTGF的蛋白表达均高于对照组(P<0.05)。Vorapaxar干预显著降低了各时间点血清TAT、PAI-1以及染尘28、56 d时大鼠血清TF水平(P<0.05),增高染尘14、28 d大鼠血清CC16的水平(P<0.05),并使染尘56 d大鼠肺组织炎症和纤维化程度减轻,肺组织Col-Ⅰ、αvβ5、αvβ6、PAR-1和CTGF蛋白表达水平降低(P<0.05)。结论 SiO_(2)吸入可导致大鼠肺上皮细胞损伤,激活凝血级联反应和增加肺组织PAR-1表达,诱发肺部炎症和纤维化;PAR-1拮抗剂可显著抑制SiO_(2)诱导的大鼠凝血级联反应,减轻肺部炎症和纤维化。Objective To explore the changes in coagulation cascade of silicosis in rats and the therapeutic effects of protease activated receptor-1(PAR-1) antagonist on the pulmonary fibrosis in rats.Methods Twenty-seven SPF grade healthy male Wistar rats were randomly divided into three groups:control,model(SiO_2),and Vorapaxar(PAR-1 antagonist) group.The silicosis rat model was established using intranasal inhalation of silica dust method(SiO_(2) concentration was 800 mg/m^(3)),6 hours/day for 28 consecutive days;the rats in Vorapaxar group were given Vorapaxar by oral from the first day of modeling.Rats were euthanized on the 14th,28th,and 56th day after silica dust exposure,collect lung tissues and serum.Then,HE and Masson's trichrome staining were used to observe the pathological and fibrotic changes in lung tissue of the rats,immunohistochemistry was performed to detect the expression of TGF-β1 in lung tissues,enzyme linked immunosorbent assay(ELISA) was used to detect the serum levels of Clara cell secretory protein-16(CC16),tissue factor(TF),thrombin antithrombin complex(TAT),and plasminogen activator inhibitor-1(PAI-1) of the rats,Western blot analysis was used to assess the protein expression of collagen typeⅠ(Col-Ⅰ),integrins αvβ5 and αvβ6,PAR-1,and connective tissue growth factor(CTGF) in lung tissues of the rats.Results The results showed that the serum levels of TF,TAT,and PAI-1 in the rats of model group were significantly increased compared to the control rats on the days of 14th,28th,and 56th after silica dust exposure(P<0.05),while the serum level of CC16 level was decreased(P<0.05);meanwhile,the degrees of inflammation and fibrosis in lungs of the rats after 56 days exposure to silica dust and the expression levels of Col-Ⅰ,αvβ5,αvβ6,PAR-1,and CTGF proteins in lung tissues were all significantly higher than that in control rats(P<0.05).Vorapaxar intervention significantly reduced serum levels of TAT and PAI-1 levels at various time points,and serum TF level in rats exposed to silica dust f

关 键 词：矽肺 纤维化 凝血级联反应 沃拉帕沙(Vorapaxar) 蛋白酶激活受体-1(PAR-1) 

分 类 号：R135.2[医药卫生—劳动卫生] R994.3[医药卫生—公共卫生与预防医学]